EP2593149B1 - Matériau composite comprenant de la pectine et du phosphate de calcium et son procédé d'obtention - Google Patents

Matériau composite comprenant de la pectine et du phosphate de calcium et son procédé d'obtention Download PDF

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Publication number
EP2593149B1
EP2593149B1 EP11749918.6A EP11749918A EP2593149B1 EP 2593149 B1 EP2593149 B1 EP 2593149B1 EP 11749918 A EP11749918 A EP 11749918A EP 2593149 B1 EP2593149 B1 EP 2593149B1
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Prior art keywords
pectin
calcium phosphate
calcium
phosphate
composite material
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German (de)
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EP2593149A2 (fr
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Paola Petrini
Maria Cristina Tanzi
Liliana Giuliano
Fabiola Munarin
Pierfrancesco Robotti
Giacomo Bianchi
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Lincotek Trento SpA
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Politecnico di Milano
Eurocoating SpA
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K33/00—Medicinal preparations containing inorganic active ingredients
    • A61K33/42—Phosphorus; Compounds thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/40—Composite materials, i.e. containing one material dispersed in a matrix of the same or different material
    • A61L27/42—Composite materials, i.e. containing one material dispersed in a matrix of the same or different material having an inorganic matrix
    • A61L27/425—Composite materials, i.e. containing one material dispersed in a matrix of the same or different material having an inorganic matrix of phosphorus containing material, e.g. apatite
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08L—COMPOSITIONS OF MACROMOLECULAR COMPOUNDS
    • C08L5/00—Compositions of polysaccharides or of their derivatives not provided for in groups C08L1/00 or C08L3/00
    • C08L5/06—Pectin; Derivatives thereof

Definitions

  • the present invention relates to a method for the realisation of a composite material comprising pectins and calcium phosphate, wherein the pectins are cross-linked by calcium phosphate; the composite material obtained by such method and the use of such composite material.
  • Pectins are hetero-polysaccharides isolated from the cell wall of plants and are widely used in the food and pharmaceutical industries.
  • Peptide polysaccharides have a very complex structure and belong to the family of polyuronic acids, such as alginate and hyaluronic acid, characterized by the presence of carboxylic groups which, depending on the pH, can be deprotonated making the polysaccharide negative.
  • the amount of carboxylic groups can vary, changing the characteristics of pectins.
  • pectins contain 1,4- ⁇ -D galacturonic acid, with a variable number of carboxyl groups present in the form of methyl-ester groups.
  • the degree of esterification (DE) is used to rank pectins into high methoxyl (HM, DE> 50) or low methoxyl (LM, DE ⁇ 50) pectins.
  • Pectins may be dissolved in water, but once cross-linked, they form water insoluble gels, with properties that depend on the type and degree of cross-linking, also depending on the degree of esterification.
  • HM pectins require very low pH for gel formation, whereas LM pectins form gels by cross-linking in the presence of divalent and trivalent cations.
  • cross-linking agents typically used are the calcium and iron chlorides, commonly used in the formation of micro- and nano-particles or coatings.
  • the kinetics of gelification determined by such compounds is very fast and therefore it is difficult to control. Similarly to what happens with alginates, this represents a significant drawback for the preparation of injectable and/or bulk gels, where homogeneity is an essential requisite.
  • organic calcium salts are better tolerated from a physiological point of view: for example, it is possible to obtain gel from calcium gluconate, although in this case, the gelification kinetics is fast compared to what happens with the calcium phosphate , as explained hereinafter.
  • calcium is released from little soluble calcium salts such as calcium carbonate, calcium citrate and calcium EDTA.
  • the release of calcium is achieved through the progressive lowering of the pH of the solution induced by the degradation of a molecule that is slowly hydrolyzed, such as ⁇ -gluconolactone.
  • These gels are prepared in conditions of lack of calcium and their mechanical properties are poor compared to those of gels obtained from solutions of calcium chloride.
  • the viscosity of a gel increases as a result of cross-linking, and depends on the amount of cross-linker and the degree of esterification of pectin.
  • the properties range from a viscous gel to solid materials. The properties are highly dependent on the molecular weight of pectin and on the pH of the environment.
  • the research is aimed mainly at identifying new substances and/or materials that improve the features of compatibility with the surrounding tissues, have good mechanical properties and a speed of resorption adjustable according to the application.
  • the materials usually applied in the medical field can be assigned to various areas.
  • injectable gels usually used based on hyaluronic acid and collagen, are used for various applications such as: lip augmentation, correction of superficial scars, fine lines and wrinkles, naso-labial folds, increases in volume, etc.
  • injectable gels are divided into materials with rapid resorption kinetics (3-6 months), medium term (6-12 months) and permanent injectable gels.
  • the materials based on hyaluronic acid may contain, as a radical scavenger, mannitol, in order, as a result of material implant, to limit the damage of the inflammatory response and increase the residence time of the same material.
  • autologous bone (autograft), homologous bone (allograft), heterologous material such as DBM (Demineralized Bone Matrix) and collagen, synthetic materials such as tricalcium phosphates and hydroxyapatite.
  • autograft autologous bone
  • homologous bone allograft
  • heterologous material such as DBM (Demineralized Bone Matrix) and collagen
  • synthetic materials such as tricalcium phosphates and hydroxyapatite.
  • these materials generally have good biological properties, such as osteogenesis but also exhibit some drawbacks.
  • autologous bone has a limited availability and very high morbidity, associated with the double operation, and phenomena of pain persistence for the patient.
  • homologous bone the risks are mainly related to the risk of transmission of infectious diseases and the techniques used to reduce this risk affect the biological properties of implants, reducing the intrinsic osteogenetic and osteoconductive potential.
  • the heterologous material does not allow a structural support, while synthetic materials have poor osteogenic properties and variable mechanical properties depending on the type of material used. 95% of these calcium phosphates are resorbed in 26 to 86 weeks.
  • the materials used in commercial products available to date are esterified hyaluronic acid and collagen, which have a proven clinical experience and good tolerability.
  • materials have been used such as polyurethanes, polycarbonates, polylactic acid and polyglycolic collagen, fibrin, chitosan optionally combined with polygalacturonic acid, silk, alginates and agarose. In particular, using chitosan it is not possible to incorporate cells within the gel matrix that is formed.
  • U.S. Patent 5980625 describes the use of a cement formed by mixing a solution of pectins with a calcium phosphate powder.
  • the cement described by such patent differs from a gel material, wherein a solution of pectin is cross-linked with a solution of calcium phosphate.
  • the mechanical properties of bone cement in fact, are very different from those of a gel and the water content is different.
  • the ratio between pectin percentages and calcium phosphate powder percentages is to the advantage of the latter.
  • DE4103876 describes the fixing of a granular bone prosthesis by mixing an aqueous solution of pectin and an aqueous solution of a calcium compound to form a glue.
  • the pectin concentration is from 0.5 to 10% and the calcium compound has a concentration of 2 to 20 %.
  • Pectin has a molecular weight of 100000 to 150000 and an esterification percentage of 5%.
  • the calcium compound is for example calcium carbonate or calcium lactate.
  • pectins Materials consisting of pectins are known, both of the high-methoxyl (HM) and low methoxyl (LM) type, and calcium phosphate blended together.
  • pectin serves as porogenic or plasticizing agent for forming materials in the form of a paste.
  • no linking reactions occur in these materials between pectin and calcium phosphate and the ratio between such components is to the benefit of calcium phosphate.
  • An object of the invention is to improve the prior art.
  • Another object of the invention is to provide a method that is simple and easy to implement.
  • a further advantage of the invention is to provide a method wherein there are only two main components.
  • Another object of the present invention is to provide a method which is inexpensive compared to known methods.
  • a further object of the present invention is to provide a material with improved mechanical and biocompatibility properties.
  • a further object of the present invention is to provide a composite material that may be obtained quickly and inexpensively.
  • a composite material is provided as specified in the independent claim 11.
  • the method object of the present invention comprises the following various steps.
  • a first step of the method object of the present invention consists in preparing a suspension of calcium phosphate at a concentration of 0.1-10% weight/volume.
  • the percentage used preferably varies between 0.25% and 5% weight/volume and more preferably between 0.25% and 3% weight/volume.
  • the calcium phosphates used consist of hydroxyapatite and/or are in other forms, non-crystalline or crystalline.
  • the following types of calcium phosphate may also be present, alone or in combination: calcium phosphate Ca3(PO4)2, calcium dihydrogen phosphate Ca(H2PO4)2, calcium hydrogen phosphate CaHPO4, tricalcium phosphate, ⁇ or ⁇ Ca3(PO4)2, tetracalcium phosphate Ca4(PO4)2, hydroxyapatite Ca10(PO4)6(OH)2.
  • calcium phosphate Other possible types are: acid calcium phosphate, calcium dihydrogen phosphate, calcium glycerophosphate, calcium hydrogen phosphate, calcium lactophosphate, tribasic calcium phosphate, calcium-o-phosphate, calcium pyrophosphate and calcium tetraphosphate.
  • the mixing of the suspension of calcium phosphates occurs for a time between 1 second and 30 minutes at room temperature.
  • the resulting solution has a neutral or slightly alkaline pH.
  • the temperature can vary from room temperature and this changes the features of the suspension.
  • the temperature in fact, varies the solubility of calcium phosphates, as well as pH and mixing time, and therefore the quantity of calcium ions in solution, which is linked to the degree of pectin cross-linking that may be obtained.
  • calcium ions released from the calcium phosphates listed above allow obtaining a composite material that may not be obtained with calcium-based substances commonly used in the prior art, such as calcium chloride.
  • the temperature can range from 0 °C to 100 °C and the mixing time can vary from 1 second to 25 hours.
  • the pH of the suspension/solution of calcium phosphates may be changed from neutral pH to acidic pH.
  • the mixing time, pH and temperature vary the percentage of calcium phosphates dissolved in the solution.
  • a second step of the method object of the present invention consists in preparing an aqueous solution of pectin present at concentrations of 0.2-10% weight/volume.
  • pectins preferably used are pectins extracted from the lemon peel and/or of different origin with a degree of esterification of between 5% and 50%, in particular pectins with a degree of esterification of 40% and/or 42%.
  • the molecular weight of these substances usually varies between 40 and 350 KDa.
  • the molecular weight of pectins varies between 15 and 1000 KDa.
  • pectins are dissolved in the aqueous solution at the percentage listed above at room temperature for 12 hours; the solution obtained has an acidic pH.
  • pectins can also be dissolved at a higher temperature: the temperature increase reduces the time of dissolution of the pectins.
  • This aqueous solution of pectins may be obtained in an alternative embodiment, at a temperature of about 80 °C for 2 hours. In a further embodiment, such solution may be obtained at 40 °C for 2 hours. More generally, the temperature may be between 0 °C and 100 °C and the mixing time may vary from 1 second to 25 hours.
  • the pH of the solution may be made neutral by the addition of bases.
  • the preparation of the pectin solution above can also be achieved with known methods generally used for pectins or other similar polysaccharides.
  • the method of the present invention can be extended to all anionic polysaccharides, similar to pectin, and all changes that may be made fall within the scope of protection of the present invention.
  • a subsequent step of the method object of the present invention consists in mixing the solution of pectin and the suspension of calcium phosphates.
  • This mixing process can take place instantaneously by mixing equal quantities by volume of the solution and suspension above at room temperature for a time ranging between 5 and 10 seconds at an acidic or neutral pH.
  • Such mixing may take place in an alternative embodiment, by gradually adding 1 part of suspension of calcium phosphates to 0.2-2 parts of pectin solution, or preferably, to 0.4-2 parts of pectin solution.
  • the temperature used in this case can be between 4 °C and 80 °C for a period of up to 30 minutes and obtaining a solution with neutral or slightly alkaline pH.
  • This method has the advantage of being able to dose the amounts and vary them in order to obtain composite materials with different features applicable to different sectors and with improved stability properties.
  • This method has the following advantages: the use of a component, such as calcium phosphates, naturally present in the human body and therefore highly biocompatible and the cross-linking reaction of pectins with calcium phosphate is slow and can therefore be controlled.
  • the composite material in the form of gel obtained from such cross-linking is homogeneous, and the cross-linking effect can be evaluated in advance.
  • the percentages of pectin and calcium phosphate are substantially equivalent or are in favor of a greater amount of pectin.
  • the structure of the composite material obtained has a high water content and mechanical and shape properties specific of substances in the form of a gel.
  • the mixture of pectin and calcium phosphates can be poured into a syringe or into a mold and let to gel for about 24 hours.
  • the gelification time may vary depending on the desired application.
  • the cross-linking of pectins takes place with at least one portion of the calcium phosphate present in solution for forming a polymeric compound with a gelatinous consistency. At least a second part of the remaining calcium phosphate, not present in solution, remains, as inorganic part, trapped inside such polymeric compound formed as a result of the cross-linking, and is a filler which gives better mechanical and reinforcing properties compared to the cross-linked polymers of the prior art, which have no filler.
  • the method according to the invention has the advantage, compared to the prior art, to obtain the cross-linking reaction of pectins starting from calcium phosphate, rather than calcium chloride that is typically used, and hence to combine in a single process step both the cross-linking of pectins and the incorporation of the inorganic part into the same.
  • the main structural components are pectins and calcium phosphate, while there may be additives or other molecules or substances which, if present, impart secondary properties to the composite material obtained by such method.
  • the advantage of speeding up the production method basically consisting of a single step, adds up to the advantage of having few essential components, making the production itself more economical.
  • a step of such method may provide for the addition of mannitol to a final concentration of mannitol between 50-500 mM.
  • the final concentration of mannitol is 220 mM.
  • This substance which is a chiral alditol, with six hydroxyl groups in the aliphatic chain consisting of six saturated carbon atoms, is a substance that can be easily found naturally in algae and fungi, and the name is derived from the manna, the sap of ash, from which it may be obtained.
  • mannitol allows increasing the stability to ionizing radiations and thermal stresses, thus preventing degradation effects related to the sterilization steps and to the duration (shelf life) of the material itself.
  • the permanence of an inorganic calcium phosphate phase in the composition is caused by the relative concentration of calcium phosphates and pectins, but also by the values of pH, temperature, time of stirring of the suspension of calcium phosphates, and type and crystallinity of the calcium phosphate used.
  • the inorganic fraction of calcium phosphate that is not dissolved depends on the features of the method according to the invention, which have effect on the same solubility of calcium phosphates.
  • the calcium phosphate particles constituting the filler, improve the biological and biocompatibility features of the composite material object of the present invention, e.g. stimulating the production of collagen or other useful substances at the site where such material is introduced.
  • the method according to the present invention may comprise the steps of loading the composite material with substances that are active towards the reduction of the infection, such as metal ions and/or topical antimicrobial drugs and/or pre-loading the calcium phosphate with metal ions known for their antibacterial effectiveness, such as, for example, silver and zinc and/or conveying, through the pectin, drugs and/or specific medicinal substances.
  • substances that are active towards the reduction of the infection such as metal ions and/or topical antimicrobial drugs and/or pre-loading the calcium phosphate with metal ions known for their antibacterial effectiveness, such as, for example, silver and zinc and/or conveying, through the pectin, drugs and/or specific medicinal substances.
  • the present invention also includes a composite material comprising pectin and calcium phosphate blended together, wherein the calcium phosphate has a concentration of between 0.1 and 10% weight/volume, and comprising pectin cross-linked with at least one portion of the calcium phosphate and at least one second portion of the calcium phosphate trapped to form an inorganic phase of calcium phosphate.
  • Pectin is present in aqueous solution at a concentration between 0.2 and 10% weight/volume.
  • Pectin has a percentage of esterification of between 5% and 50% or 42% or 40%.
  • the pectins according to the present invention have a molecular weight between 40 and 350 kDa, or between 15 and 1000 kDa.
  • Calcium phosphate is present in aqueous suspension at a concentration of between 0.25% and 5% weight/volume or between 0.25% and 3%.
  • Such calcium phosphate comprises one or more of the following types: calcium phosphate Ca3(PO4)2, calcium dihydrogen phosphate Ca(H2PO4)2, calcium hydrogen phosphate CaHPO4, tricalcium phosphate ⁇ or ⁇ Ca3(PO4)2, tetracalcium phosphate Ca4(PO4)2, hydroxyapatite Ca10(PO4)6(OH)2, acid calcium phosphate, calcium glycerophosphate, calcium lactophosphate, tribasic calcium phosphate, calcium-o-phosphate and calcium pyrophosphate.
  • the composite material according to the present invention is a hydrogel. It includes equal parts of the pectin solution and the suspension of calcium phosphate mixed together or it comprises from 0.2 to 2 parts of the solution of pectin, or preferably 0.4 to 2 parts of the solution of pectin, and a part of the suspension of calcium phosphate.
  • the composite material according to the present invention further includes mannitol.
  • Mannitol has a final concentration of between 50 and 500 mM or 220 mM.
  • the composite material according to the present invention is obtained for use in medical applications as a medicine for aesthetic applications and/or the treatment of skin lesions and/or for the treatment of cartilage defects and/or the bone substitution and/or for similar applications.
  • the composite material according to the present invention can be loaded with active substances in relation to the reduction of the infection, such as metal ions and/or topical antimicrobial drugs and/or calcium phosphate may be pre-loaded with metal ions known for their antibacterial effectiveness such as, for example, silver and zinc and/or the pectin may be suitable for conveying drugs and/or specific medicinal substances.
  • active substances such as metal ions and/or topical antimicrobial drugs and/or calcium phosphate
  • metal ions known for their antibacterial effectiveness such as, for example, silver and zinc
  • the pectin may be suitable for conveying drugs and/or specific medicinal substances.
  • Examples 1-6 below, and the respective tables 1-9, show some embodiments of the method according to the invention for obtaining a composite material with specific features. Based on the materials obtained according to examples 1 and 2, stability tests, analysis to evaluate the total content of water, tests of swelling in water, as shown in figure 1 , infrared spectroscopy, as shown in figures 2 and 3 , analysis of injectability and extrudability of the composite material, etc., have subsequently been performed.
  • This composite material may be used with various applications in the medical field.
  • such composite material present in the form of injectable gel, may be used for aesthetic applications, such as: lip augmentation, correction of superficial scars, fine lines and wrinkles, naso-labial folds, increases in volume, etc.
  • An example of material used for such applications can be represented by table 3 and/or may occur according to other parameters described in the examples herein. Additionally, such composite material may also be applied in the treatment of skin lesions.
  • a composite material according to the invention, modulated in order to obtain a membrane or a three-dimensional support should possess, in addition to the advantages already mentioned for the previous applications (biological, commercial and production advantages), an advantage related to the possibility of loading such material with active substances in relation to the reduction of the infection, such as metal ions, or topical antimicrobial drugs. Further possibilities presented by such composite material are: pre-loading the calcium phosphate contained therein with metal ions known for their antibacterial effectiveness, such as, for example, silver and zinc; using the solution of pectin as a carrier for specific drugs.
  • microspheres suitable for drug delivery that could be incorporated in the bandage material used in the treatment of such skin lesions.
  • the composite material used for this purpose has the following features: high biocompatibility, absence of induced allergic phenomena, maintaining high humidity at the interface, capability of absorbing excess exudate and providing proper thermal insulation, possibility of removal without trauma, high mechanical resistance also at the hydrated state, selective permeability to gases but not to pathogens.
  • the composite material may also be used for the treatment of cartilage defects, such as a three-dimensional support, presenting the following advantages: biological advantages (no human-derivatives, low or no risk of allergies, degradability modulated depending on the application, higher stability compared to hyaluronic acid) and productivity advantages (composite material prepared by a single production step).
  • biological advantages no human-derivatives, low or no risk of allergies, degradability modulated depending on the application, higher stability compared to hyaluronic acid
  • productivity advantages composite material prepared by a single production step.
  • the use of a composite material can also result in improved mechanical performance compared to single-component materials.
  • the composite material used for this purpose has the following features: biocompatibility, excellent cellular adhesion and three-dimensional structure suitable for cellular proliferation, high porosity, gradual bio-resorption without the release of acids and with stimulation of the formation of natural cartilage tissue, high mechanical properties.
  • This composite material may also be used as an injectable gel.
  • injectable gel may be loaded with cellular components that are embedded within the matrix itself, to allow a therapy usable by minimally invasive surgery in a single step.
  • the injectable gel may also be formulated to allow a cross-linking in situ, allowing the entrapment of the preloaded cellular components and creating an environment suitable for the maintenance of cellular viability and formation of extracellular matrix, optimal conditions for the care of these cartilage defects.
  • this composite material is also possible to obtain bone substitutes to use in dentistry to cure periodontal defects, extra-alveolar defects, bone lesions, cystic cavities, intrabony defects, the maxillary sinus lift, the increase of the alveolar bridge, endodontic perforations, post-extraction sites, etc..; in orthopedics for treating fractures of the tibial head, distal radius, heel, proximal and distal tibia, proximal and distal femur, generic metaphysis, proximal humerus, acetabulum, tibial plateau, spine, bone cysts, autologous bone sampling sites, pseudoarthrosis, spinal fusion, generic bone defects, total hip revision, foot and hand surgery, arthroplasty, trauma, maxillofacial surgery, surgery of the skull, bone defects caused by cancer, etc.
  • the advantages of the composite material as a bone substitute are: as much as possible unlimited availability, rapid conversion into bone matrix, precise possibility of modulating the morphology, stability in situ, reproducible biological and mechanical features and immediate compression stability for the bone substitutes designed to withstand loads.
  • Another possible application of the composite material obtained by the present invention can envisage loading the same with cellular components, drugs, peptides or proteins or antibacterial agents, to form a controlled release vehicle of such active principles or of such cellular components.
  • Such composite material may also be used as injectable gel for applications not subject to load and be applied as a binder for using ceramic material and as filler for small bone defects such as periodontal defects.
  • Table 1 below shows the different concentrations of pectin and calcium phosphate used and the stability in water and in NaCl 0.9% w/v at at least 24 hours of the resulting composite material in gel.
  • Table 1 Concentrations of pectin and calcium phosphate, stability in water and in NaCl 0.9% w/v at 37 °C Pectin (% w/v) Calcium phosphate (% w/v) Stability in water Stability in NaCl 0.9% w/v 2 0,5 stable instable 1 stable - 2 stable - 3 stable instable 3 0,5 stable stable up to 60 min 1 - - 2 - - 3 stable stable up to 30 min 4 0,5 stable stable up to 180 min 1 - - 2 - - 3 stable stable up to 180 min
  • the composite gel materials at 2% pectin were lyophilized and water swelling tests were conducted thereon at 37 °C, as shown in Figure 1 .
  • the percentage of swelling in water was proportional to the content of calcium phosphate and, therefore, to the degree of cross-linking of the composite material.
  • the composite freeze-dried gels materials described above were also characterized by infrared spectroscopy (FT-IR) and compared with the spectra of a gel obtained by cross-linking pectin with CaCl 2 .
  • FT-IR infrared spectroscopy
  • Table 2 shows the various concentrations of pectin and calcium phosphate used and the stability in water and in NaCl 0.9% w/v at at least 24 hours of the resulting gel composite materials. The total content of water for all of such materials proved to be more than 98% by weight.
  • the gels are prepared according to table 3.
  • a pectin with a molecular weight below 200 KDa and esterification below 35% is dissolved at room temperature to a final concentration of 6% weight/volume.
  • the pH of the solution is brought to 5 with NaOH 0.5 M.
  • HA hydroxyapatite
  • the rheological properties of samples obtained were evaluated 7 days after preparation and storage at 4 °C of the same.
  • Table 9 below describes the samples produced according to examples 4 and 5.
  • Table 9 Sample Concentration of HA (weight/volume) Ratio between pectin solution and volume of the suspension of HA P4H03 (1) 0.9 1:0.5 P3H05 (2) 1.0 1:1 P3H03 (3) 0.6 1:1 P2H05 (4) 0.75 1:2 P2H03 (5) 0.45 1:2
  • the gel is prepared by following the method and the components listed in table 4.
  • the pectin with molecular weight between 500 and 600 KDA and esterification of 37% was dissolved at room temperature to a final concentration of 6% weight/volume.
  • the pH of the solution was brought to 5 with NaOH 0.5 M.
  • HA hydroxyapatite
  • the gels were prepared as in example 5. Mannitol was added to the samples at a final concentration of 220 mM mannitol. These samples were obtained by dissolving mannitol in the solution of pectin. The gels were then subjected to beta radiation at 25 KGy and subsequently analyzed by the rheological tests. The storage module and the complex viscosity of each gel were quantified according to rheological measurements carried out as described in example 4. These measurements are shown in figures 11 and 12 .

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Claims (19)

  1. Procédé pour la réalisation d'un matériau composite comprenant des pectines et du phosphate de calcium, comprenant les étapes suivantes:
    - la préparation d'une solution aqueuse de pectine,
    - la préparation d'une suspension aqueuse de phosphate de calcium,
    - le mélange de ladite solution de pectine et ladite suspension aqueuse de phosphate de calcium,
    dans lequel ladite étape de préparation de la suspension aqueuse de phosphate de calcium est effectuée avec une concentration de phosphate de calcium comprise entre 0,1 et 10% en poids/volume, caractérisé en ce qu'il comprend une étape de réticulation de ladite solution de pectine avec au moins une portion du phosphate de calcium dissous dans ladite suspension aqueuse, en piégeant ainsi au moins une deuxième portion dudit phosphate de calcium en formant une phase inorganique du phosphate de calcium.
  2. Procédé selon la revendication 1, dans lequel ladite étape de préparation d'une solution aqueuse de pectine est effectuée en utilisant une concentration de pectine comprise entre 0,2 et 10% en poids/volume.
  3. Procédé selon la revendication 1 ou 2, dans lequel ladite étape de préparation d'une solution aqueuse de pectine est effectuée avec de la pectine ayant un pourcentage d'estérification compris entre 5% et 50% ou de 42% ou 40% et/ou dans lequel ladite étape de préparation d'une solution aqueuse de pectine est effectuée avec de la pectine ayant un poids moléculaire compris entre 15 et 1000 kDa ou ayant un poids moléculaire compris entre 40 et 350 kDa.
  4. Procédé selon une des revendications précédentes, dans lequel ladite étape de préparation d'une solution aqueuse de pectine est réalisée par mélange effectué à une température entre 0 C et 100 C pendant un temps de mélange entre 1 seconde et 25 heures ou est réalisée par mélange effectué à une température de 40 C pendant un temps de mélange de 2 heures.
  5. Procédé selon la revendication 1, dans lequel ladite préparation de ladite suspension aqueuse de phosphate de calcium est réalisée avec une concentration comprise entre 0,25% et 5% en poids/volume ou entre 0,25% et 3% en poids/volume.
  6. Procédé selon l'une quelconque des revendications précédentes, dans lequel ladite étape de préparation d'une suspension aqueuse de phosphate de calcium est réalisée avec un ou plusieurs des types suivants de phosphate de calcium: phosphate de calcium Ca3(PO4)2, dihydrogénophosphate de calcium Ca(H2PO4)2, hydrogénophosphate de calcium CaHPO4, phosphate tricalcique β ou α Ca3(PO4)2, phosphate tétracalcique Ca4(PO4)2, hydroxyapatite Ca10(PO4)6(OH)2, phosphate de calcium acide, glycérophosphate de calcium, lactophosphate de calcium, phosphate de calcium tribasique, phosphate-o-calcium et pyrophosphate de calcium.
  7. Procédé selon l'une quelconque des revendications précédentes, dans lequel ladite étape de préparation d'une suspension aqueuse de phosphate de calcium comprend les étapes suivantes:
    - mélange à un pH neutre ou un pH acide ou un pH alcalin et/ou
    - à des températures entre 0°C et 100°C et/ou
    - pendant un temps de mélange entre 1 seconde et 25 heures.
  8. Procédé selon la revendication 1, dans lequel ladite étape de mélange de la solution de pectine et de la suspension aqueuse de phosphate de calcium est réalisée en mélangeant des parties égales de la solution de pectine et de la suspension aqueuse de phosphate de calcium ou une partie de la suspension aqueuse de phosphate de calcium avec une fraction de solution de pectine comprise entre 0,2 parties et 2 parties ou entre 0,4 parties et 2 parties.
  9. Procédé selon l'une quelconque des revendications précédentes, comprenant une étape d'ajout de mannitol ou comprenant une étape d'ajout de mannitol à ladite solution de pectine et/ou dans lequel ladite étape d'ajout est effectuée en obtenant une concentration finale de mannitol comprise entre 50 and 500 mM or en obtenant une concentration finale de mannitol de 220 mM.
  10. Procédé selon l'une quelconque des revendications précédentes, comprenant les étapes suivantes:
    - le chargement dudit matériau composite avec des substances actives contre la réduction de l'infection, des ions métalliques et/ou des médicaments antimicrobiens topiques et/ou
    - le pré-chargement dudit phosphate de calcium avec des ions métalliques connus pour leur efficacité antibactérienne, argent ou zinc et/ou
    - le transport, à travers ladite pectine, de médicaments et/ou de substances médicinales spécifiques.
  11. Matériau composite sous la forme d'hydrogel comprenant de la pectine et du phosphate de calcium mélangés ensemble, dans lequel ledit phosphate de calcium a une concentration comprise entre 0,1 et 10% en poids/volume, caractérisé en ce qu'il comprend ladite pectine réticulée avec au moins une portion de phosphate de calcium et au moins une deuxième portion de phosphate de calcium piégée pour former une phase inorganique de phosphate de calcium.
  12. Matériau composite selon la revendication 11, dans lequel ladite pectine est présente en solution aqueuse avec une concentration entre 0,2 et 10% en poids/volume et/ou dans lequel ladite pectine a un pourcentage d'estérification compris entre 5% et 50% ou de 42% ou 40%.
  13. Matériau composite selon l'une quelconque des revendications 11 ou 12, dans lequel ladite pectine a un poids moléculaire entre 40 et 350 kDa ou un poids moléculaire entre 15 et 1000 kDa.
  14. Matériau composite selon une des revendications 11 à 13, dans lequel ledit phosphate de calcium est présent en suspension aqueuse avec une concentration comprise entre 0,25 et 5% en poids/volume ou avec une concentration comprise entre 0,25% et 3%.
  15. Matériau composite selon une des revendications 11 à 14, dans lequel ledit phosphate de calcium comprend un ou plusieurs des types suivants: phosphate de calcium Ca3(PO4)2, dihydrogénophosphate de calcium Ca(H2PO4)2, hydrogénophosphate de calcium CaHPO4, phosphate tricalcique β ou α Ca3(PO4)2, phosphate tétracalcique Ca4(PO4)2, hydroxyapatite Ca10(PO4)6(OH)2, phosphate de calcium acide, glycérophosphate de calcium, lactophosphate de calcium, phosphate de calcium tribasique, phosphate-o-calcium et pyrophosphate de calcium.
  16. Matériau composite selon une des revendications 11 à 15, dans lequel ledit matériau composite comprend une portion égale de la solution de pectine et de ladite suspension aqueuse de phosphate de calcium mélangées ensemble ou dans lequel ledit matériau composite comprend de 0,2 parties à 2 parties de ladite solution de pectine, ou de 0,4 à 2 parties de ladite solution de pectine, et une partie de ladite suspension aqueuse de phosphate de calcium.
  17. Matériau composite selon l'une quelconque des revendications 11 à 16, comprenant du mannitol et/ou dans lequel ledit mannitol a une concentration finale comprise entre 50 et 500 mM ou de 220 mM.
  18. Matériau composite selon l'une quelconque des revendications 11 à 17, dans lequel ledit matériau est chargé avec des substances actives contre la réduction de l'infection, des ions métalliques et/ou des médicaments antimicrobiens topiques et/ou dans lequel ledit phosphate de calcium est pré-chargé avec des ions métalliques connus pour leur efficacité antibactérienne, argent ou zinc et/ou dans lequel ladite pectine est adaptée pour transporter des médicaments et/ou des substances médicinales spécifiques.
  19. Matériau composite selon l'une quelconque des revendications 11 à 18, adapté pour l'utilisation dans des applications médicales comme un médicament pour des applications esthétiques et/ou le traitement de lésions de la peau et/ou pour le traitement de défauts du cartilage et/ou comme substitut osseux.
EP11749918.6A 2010-07-13 2011-07-13 Matériau composite comprenant de la pectine et du phosphate de calcium et son procédé d'obtention Active EP2593149B1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
IT000140A ITVR20100140A1 (it) 2010-07-13 2010-07-13 Materiale composito formato da pectina e da calcio fosfato e metodo per la sua realizzazione
PCT/IB2011/053136 WO2012007917A2 (fr) 2010-07-13 2011-07-13 Matériau composite comprenant de la pectine et du phosphate de calcium et son procédé d'obtention

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EP2593149B1 true EP2593149B1 (fr) 2014-04-30

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ITVR20120045A1 (it) 2012-03-14 2013-09-15 Eurocoating S P A Metodo per la realizzazione di un impianto osseo biocompatibile comprendente elementi granulari ed un gel biopolimerico bioriassorbibile e impianto oseeo biocompatibile ottenuto con tale metodo.
CN114538914A (zh) * 2022-03-14 2022-05-27 成都理工大学 一种具有抗菌功能的锌掺杂磷酸钙陶瓷微球及制备方法

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DE2708404A1 (de) * 1977-02-26 1978-08-31 Herbstreith Pektinfab Verfahren zur herstellung eines pektinderivates, insbesondere auf der basis von obstpektin, und zur herstellung eines nahrungsmittel-gelierstoffes und nahrungsmittel-gelierstoff
DE4103876C2 (de) * 1990-02-09 1994-07-07 Asahi Optical Co Ltd Verfahren zur Fixierung einer körnigen Knochen-Prothese und Kit zur Herstellung einer körnigen Knochen-Prothesezusammensetzung zur Verwendung in dem Fixierungsverfahren
JPH10248915A (ja) * 1997-03-13 1998-09-22 Ngk Spark Plug Co Ltd リン酸カルシウムセメント硬化液及びリン酸カルシウムセメント硬化体用組成物
US7494669B2 (en) * 2001-02-28 2009-02-24 Carrington Laboratories, Inc. Delivery of physiological agents with in-situ gels comprising anionic polysaccharides
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WO2012007917A2 (fr) 2012-01-19
BR112013000896B1 (pt) 2019-04-24
US20130177656A1 (en) 2013-07-11
BR112013000896A2 (pt) 2016-05-24
WO2012007917A3 (fr) 2012-04-12
EP2593149A2 (fr) 2013-05-22
US9066934B2 (en) 2015-06-30
ITVR20100140A1 (it) 2012-01-14
BR112013000896B8 (pt) 2023-02-28

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